Figure 6. Role of L-plastin in NKG2D-mediated cell migration.

(A) Human NK cells were transfected with siRNA targeting L-plastin or a siRNA control. L-plastin expression levels were analyzed by Western blot, and anti-GAPDH blots were included as protein-loading controls. (B) Twenty-four hours after transfection, cells were incubated with anti-NKG2D or IgG, and their ability to migrate toward a CXCL12 gradient was assayed. Data shown are the mean and sem of two replicates pooled from three independent experiments. The effect of L-plastin depletion upon activation was assessed by ANOVA (P=0.025). (C) F-actin content was measured in human NK cells transfected with siRNA for L-plastin or a siRNA control. After activations, cells were fixed, permeabilized, and stained with phalloidin-TRITC, and the mean fluorescence intensity was determined. Results indicate the mean and sem of three independent experiments. Data were examined by ANOVA. Statistical significance for the effect of L-plastin depletion upon receptor activation on F-actin content was P=0.004.